An Anaerobic Environment Drives the Harboring of Helicobacter pylori within Candida Yeast Cells (original) (raw)
Related papers
Biology, 2021
Helicobacter pylori is capable of entering into yeast, but the factors driving this endosymbiosis remain unknown. This work aimed to determine if temperatures outside the optimal range for H. pylori increase its harboring within Candida. H. pylori strains were co-cultured with Candida strains in Brucella broth supplemented with 5% fetal bovine serum and incubated at 4, 25, 37 or 40 °C. After co-culturing, yeasts containing bacteria-like bodies (Y-BLBs) were observed by optical microscopy, and the bacterium were identified as H. pylori by FISH. The H. pylori 16S rRNA gene was amplified from the total DNA of Y-BLBs. The viability of intra-yeast H. pylori cells was confirmed using a viability assay. All H. pylori strains were capable of entering into all Candida strains assayed. The higher percentages of Y-BLBs are obtained at 40 °C with any of the Candida strains. H pylori also increased its harboring within yeast in co-cultures incubated at 25 °C when compared to those incubated at 3...
Nutrient Deficiency Promotes the Entry of Helicobacter pylori Cells into Candida Yeast Cells
Biology
Helicobacter pylori, a Gram-negative bacterium, has as a natural niche the human gastric epithelium. This pathogen has been reported to enter into Candida yeast cells; however, factors triggering this endosymbiotic relationship remain unknown. The aim of this work was to evaluate in vitro if variations in nutrient concentration in the cultured medium trigger the internalization of H. pylori within Candida cells. We used H. pylori–Candida co-cultures in Brucella broth supplemented with 1%, 5% or 20% fetal bovine serum or in saline solution. Intra-yeast bacteria-like bodies (BLBs) were observed using optical microscopy, while intra-yeast BLBs were identified as H. pylori using FISH and PCR techniques. Intra-yeast H. pylori (BLBs) viability was confirmed using the LIVE/DEAD BacLight Bacterial Viability kit. Intra-yeast H. pylori was present in all combinations of bacteria–yeast strains co-cultured. However, the percentages of yeast cells harboring bacteria (Y-BLBs) varied according to ...
Foodborne Yeasts Serve as Reservoirs of Helicobacter Pylori
Journal of Food Safety, 2012
It is not clear where Helicobacter pylori establishes in the environment and how it is transmitted to humans. Fifteen yeasts were isolated from 15 food samples. Microscopic examination of yeasts revealed fast-moving bacterium-like bodies (BLBs) inside the vacuoles of yeasts. Yeasts were identified as Candida species. Polymerase chain reaction (PCR) was recruited to detect H. pylori-specific ureAB (urease) and babAB (BabA/B adhesins) genes in the total DNAs of yeasts. The ureAB gene was amplified from 9/15 yeasts and babAB from 2/15. Sequencing of the PCR products of ureAB (406 bp) and babAB (980 bp) genes that were amplified from one yoghurt yeast showed 98% homology with those of control H. pylori. Fluorescent microscopy of stained yeasts showed live BLBs inside the vacuoles. It is proposed that foodborne yeast which originates from the environment could accommodate H. pylori in nature and act as a vehicle for its transmission to the human gastrointestinal tract.
Immunodetection of Helicobacter pylori-specific Proteins in Oral and Gastric Candida Yeasts
Archives of Iranian Medicine, 2013
BACKGROUND: Human gastric epithelium and immunocytes have been recognized as the sole specialized eukaryotic cells that host Helicobacter pylori (H. pylori). The aim of this study was to provide further evidence for our previous proposal regarding the occurrence of H.pylori inside the yeast vacuole, verifying the viability of the intravacuolar H.pylori by western blotting.METHODS: Light microscopy and polymerase chain reaction (PCR) were used for primary detection of nonculturable H.pylori in 11 Candida yeasts (six oral and five gastric). Boiling was used for extraction of proteins from yeasts and the control H.pylori. Western blot analysis was recruited to assess the occurrence of H.pylori-specific proteins in protein pool of yeasts, using IgY-Hp raised in hens and IgG1-Hp raised in mice.RESULTS: The fast-moving bacterium-like bodies (BLBs) were identified as H.pylori by amplification of H.pylori 16S rRNA, ureAB, vacA s1, and ahpC genes from the whole DNA of yeasts. Analysis of the sequenced products of 16S rRNA gene amplified from the yeast and H.pylori isolates of patient #2 showed 100 % homology with the corresponding sequences of the reference H. pylori strains in GenBank. According to published data, it was plausible to assign the H.pylori-specific proteins, detected by western blot analysis, as thiol peroxidase (21 kDa), peroxiredoxin (AhpC) (26 kDa), urease-A subunit (UreA) (32 kDa), vacuolating cytotoxin A (VacA) small subunit (36 kDa), and VacA large subunit (56 kDa).CONCLUSION: Results of this study show that inside yeast, H.pylori expresses proteins and is viable. These proteins appear to serve as powerful tools to help H.pylori to establish in the vacuole of yeast where it can reach nutrients and multiply. The intimate relationship between H.pylori and Candida yeast which began long time ago, could have led to the establishment of H.pylori inside the yeast vacuole before invading human cells.
In Vitro Incorporation of Helicobacter pylori into Candida albicans Caused by Acidic pH Stress
Pathogens, 2020
Yeasts can adapt to a wide range of pH fluctuations (2 to 10), while Helicobacter pylori, a facultative intracellular bacterium, can adapt to a range from pH 6 to 8. This work analyzed if H. pylori J99 can protect itself from acidic pH by entering into Candida albicans ATCC 90028. Growth curves were determined for H. pylori and C. albicans at pH 3, 4, and 7. Both microorganisms were co-incubated at the same pH values, and the presence of intra-yeast bacteria was evaluated. Intra-yeast bacteria-like bodies were detected using wet mounting, and intra-yeast binding of anti-H. pylori antibodies was detected using immunofluorescence. The presence of the H. pylori rDNA 16S gene in total DNA from yeasts was demonstrated after PCR amplification. H. pylori showed larger death percentages at pH 3 and 4 than at pH 7. On the contrary, the viability of the yeast was not affected by any of the pHs evaluated. H. pylori entered into C. albicans at all the pH values assayed but to a greater extent a...
Vacuoles of Candida yeast as a specialized niche forHelicobacter pylori
World Journal of Gastroenterology, 2014
Helicobacter pylori (H. pylori) are resistant to hostile gastric environments and antibiotic therapy, reflecting the possibility that they are protected by an ecological niche, such as inside the vacuoles of human epithelial and immune cells. Candida yeast may also provide such an alternative niche, as fluorescently labeled H. pylori were observed as fast-moving and viable bacterium-like bodies inside the vacuoles of gastric, oral, vaginal and foodborne Candida yeasts. In addition, H. pylori-specific genes and proteins were detected in samples extracted from these yeasts. The H. pylori present within these yeasts produce peroxiredoxin and thiol peroxidase, providing the ability to detoxify oxygen metabolites formed in immune cells. Furthermore, these bacteria produce urease and VacA, two virulence determinants of H. pylori that influence phago-lysosome fusion and bacterial survival in macrophages. Microscopic observations of H. pylori cells in new generations of yeasts along with amplification of H. pylori-specific genes from consecutive generations indicate that new yeasts can inherit the intracellular H. pylori as part of their vacuolar content. Accordingly, it is proposed that yeast vacuoles serve as a sophisticated niche that protects H. pylori against the WJG 20 th Anniversary Special Issues (6): Helicobacter pylori TOPIC HIGHLIGHT environmental stresses and provides essential nutrients, including ergosterol, for its growth and multiplication. This intracellular establishment inside the yeast vacuole likely occurred long ago, leading to the adaptation of H. pylori to persist in phagocytic cells. The presence of these bacteria within yeasts, including foodborne yeasts, along with the vertical transmission of yeasts from mother to neonate, provide explanations for the persistence and propagation of H. pylori in the human population. This Topic Highlight reviews and discusses recent evidence regarding the evolutionary adaptation of H. pylori to thrive in host cell vacuoles.
Middle East Journal of Digestive Diseases, 2020
BACKGROUND Sugar-rich foods are of the main components of daily human meals. These foods with high sugar and low water content kill bacteria. However, osmotolerant yeasts survive and multiply. The aim of this study was to examine the occurrence of intracellular Helicobacter pylori (H. pylori) and Staphylococcus spp. in yeast isolates from sugar-rich foods. METHODS Thirty-two yeast isolates from fresh fruits, dried fruits, commercial foods, and miscellaneous foods were identified by the sequencing of amplified products of 26S rDNA. Fluorescence microscopy and LIVE/DEAD bacterial viability kit were used to examine the occurrence of live bacteria inside the yeast’s vacuole. Immunofluorescence assay was used to confirm the identity of intracellular bacteria as H. pylori and Staphylococcus . Polymerase chain reaction (PCR) was used for the detection of 16S rDNA of H. pylori and Staphylococcus in the total DNA of yeasts. RESULTS Yeasts were identified as members of seven genera; Candida, ...
Yeast of the oral cavity is the reservoir of Heliobacter pylori
Journal of Oral Pathology & Medicine, 2008
BACKGROUND: Frequent occurrence of Helicobacter pylori in the human gastrointestinal tract and its persistence due to unsuccessful antimicrobial therapy might be related to a stage in the life cycle of H. pylori in which the bacterium establishes itself as an intracellular symbiont in yeast. In this study, occurrence of non-culturable H. pylori in the oral yeast was assessed by targeting vacuolating cytotoxin A (vacA s1s2) and ureAB genes in the total DNAs of yeasts. METHODS: DNAs were extracted from 13 oral yeasts in which bacterium-like bodies, suspected to be H. pylori, were observed microscopically. Primers were recruited to amplify vacA s1s2 and ureAB genes. DNAs from H. pylori and E. coli were used as controls. The amplicons from one yeast and H. pylori were sequenced. Yeasts were identified as Candida albicans. RESULTS: Fragments of vacA s1s2 and ureAB genes were amplified from 13 yeasts. The size of PCR products was 286 bp for vacA s1s2 gene and 406 bp for ureAB gene. Similar bands were obtained from the control H. pylori, and the results for E. coli were negative. The data from sequencing of PCR products showed about 98% homology between the genes amplified from yeast and those from H. pylori. CONCLUSIONS: The results of this study showed the intracellular occurrence of H. pylori in yeast. This endosymbiotic relationship might explain the persistence of H. pylori in the oral cavity, the consequence of which could be reinoculation of the stomach by the bacterium and spread of infection among human populations.
Pathogens, 2021
First-line treatment for Helicobacter pylori includes amoxicillin and clarithromycin or metronidazole plus a proton pump inhibitor. Treatment failure is associated with antibiotic resistance and possibly also with internalization of H. pylori into eukaryotic cells, such as yeasts. Factors triggering the entry of H. pylori into yeast are poorly understood. Therefore, the aim of this study was to evaluate whether clarithromycin or amoxicillin trigger the entry of H. pylori into C. albicans cells. Methods: H. pylori J99 and C. albicans ATCC 10231 were co-cultured in the presence of subinhibitory concentrations of amoxicillin and clarithromycin as stressors. Bacterial-bearing yeasts were observed by fresh examination. The viability of bacteria within yeasts was evaluated, confirming the entry of bacteria into Candida, amplifying, by PCR, the H. pylori16S rRNA gene in total yeast DNA. Results: Amoxicillin significantly increased the entry of H. pylori into C. albicans compared to the con...
Localization of H.pylori within the Vacuole of Candida Yeast by Di- UHFW,PPXQR¿XRUHVFHQFH7HFKQLTXH
2013
AbstractBackground: Reports indicate that H.pylori is able to invade the eukaryotic cells and establish inside their vacuoles. In this study, FITC-conjugated IgY-Hp was used to localize H.pylori inside the vacuole of Candida yeast. Presence of intracellular H.pylori inside the new generations of yeast cells was also examined by light microscopy and Live/Dead Bac Light staining method. Methods: $VLQJOHFRORQ\RIIUHVK\HDVWFXOWXUHZDVFXOWLYDWHGLQDOPHGLXPFRQWDLQLQJ\HDVWH[WUDFWDQG1DFHW\OJOXFRVHDPLQHVXSSOHPHQWHGZLWKIHWDOERYLQHVHUXP$IWHUKULQFXEDWLRQDW),7&FRQMXJDWHG,J<+SZDVDGGHG$IWHUKRXUV/RI\HDVWVXVSHQVLRQZDVVPHDUHGRQDJODVVVOLGHDLUGULHGDQGH[DPLQHGE\AXRUHVFHQWPLFURVFRS\:HWPRXQWVRI\HDVWFXOWXUHDQG/LYH'HDG Bac /LJKWVWDLQHGSUHSDUDWLRQVZHUHH[DPLQHGE\OLJKWDQGAXRUHVFHQWPLFURVFRS\UHVSHFWLYHO\3KRWRJUDSKVZHUHWDNHQIURPWKHIDVWmoving H.pylori inside the yeast vacuoles. Results: )OXRUHVFHQWPLFURVF...